Antisense basic fibroblast growth factor gene transfer reduces early intimal thickening in a rabbit femoral artery balloon injury model.
OBJECTIVE
The purpose of this study was to determine whether endogenous basic fibroblast growth factor (bFGF) production contributes to the intimal hyperplastic response to injury in a model of rabbit femoral artery balloon injury. Inhibition of de novo production of bFGF protein was targeted by intramural adenoviral gene transfer of antisense bFGF (Ad.ASbFGF) RNA. The adenovirus was delivered locally intraluminally at the time of arterial injury.


METHODS
New Zealand White rabbits underwent balloon injury of the superficial femoral artery, followed by intraluminal delivery of an adenoviral vector encoding a rat antisense bFGF (ASbFGF) transcript at a concentration of 1 x 10(10) plaque-forming units per milliliter. Control animals were treated in a similar fashion, using either an adenovirus encoding the lac Z reporter gene (Ad.lacZ) or phosphate-buffered saline solution (PBS; vehicle) alone. Two weeks after balloon injury, rabbits were killed and perfusion fixed. Femoral arteries were harvested for histomorphometric analysis. Intimal and medial wall thickness was measured at eight points around the vessel perimeter, and mean intimal/medial (I/M) thickness ratios were compared by analysis of variance and Student's t test. In addition, medial cell proliferation in Ad.ASbFGF and Ad.lacZ treated arteries was evaluated 4 days and 2 weeks after balloon injury by 5-Bromo-2'-deoxyuridine labeling.


RESULTS
At 14 days (n = 25) after balloon injury, histomorphometric analysis revealed a significant inhibition of intimal thickening in Ad.ASbFGF-treated arteries as compared with Ad.lacZ-treated and PBS-treated controls (I/M thickness ratios +/- SD, 0.43 +/- 0.22 for Ad.ASbFGF vs 1.03 +/- 0.28 for Ad.lacZ and 0.86 +/- 0.19 for PBS; p < 0.0001 and p = 0.004, respectively). There was no significant difference in the I/M thickness ratios of Ad.lacZ-treated and PBS-treated vessels (p = 0.27). Although there was no significant difference in the proliferation index of Ad.ASbFGF-treated and Ad.lacZ-treated vessels 4 days after injury, an increase in apoptosis was noted in the Ad.ASbFGF-treated vessels 4 days after balloon injury.


CONCLUSIONS
The use of ASbFGF RNA gene transfer, designed to inhibit de novo bFGF synthesis after balloon injury, results in a significant inhibition of neointimal formation. This suggests that continued bFGF synthesis contributes to the arterial response to injury in rabbits. ASbFGF gene transfer may be an effective strategy in limiting the intimal hyperplastic response after vascular reconstructive procedures.
Neschis
D G
DG
Department of Surgery, University of Pennsylvania Medical Center, Philadelphia, USA.
Safford
S D
SD
Hanna
A K
AK
Fox
J C
JC
Golden
M A
MA
D000256:Adenoviridae; D000818:Animals; D016375:Antisense Elements (Genetics) / Q000235:genetics*; D002404:Catheterization; D002455:Cell Division; D005263:Femoral Artery / Q000473:pathology*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000502:physiology*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006965:Hyperplasia; D007763:Lac Operon; D011817:Rabbits; D017539:Tunica Intima / Q000473:pathology*; D017540:Tunica Media / Q000473:pathology
D016375:Antisense Elements (Genetics); D016222:Fibroblast Growth Factor 2